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MB Sample ID: SA051037
| Local Sample ID: | 141215bctsa24_1 |
| Subject ID: | SU000912 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Fifth-generation C57/BL6 Pkd1tm2Ggg (Piontek et al., 2004) mice were crossed to the reporter mice C57/BL6 congenic B6.129S4-Gt(ROSA)26Sortm1Sor/J (stock 003474, Jackson Laboratories) and to C57/BL6 tamoxifen-Cre (B6.Cg-Tg(Cre/Esr1)5Amc/J mice (stock 004682), Jackson Laboratories) or Ksp-Cre B6.Cg-Tg(Cdh16-cre)91Igr/J(stock 012237), Jackson Laboratories). |
| Gender: | Males |
| Animal Animal Supplier: | Jackson Laboratories |
| Species Group: | Mammal |
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Collection:
| Collection ID: | CO000906 |
| Collection Summary: | Kidneys from two Pkd1cko/cko animals (121112-C: male, bP12; and 94414: male, P463) were harvested, minced and digested using a collagenase/hyaluronidase solution (Stemcell technologies, cat. no. 07912) followed by proximal or collecting/distal tubule cell enrichment using, respectively, biotinylated Lotus tetragonolobus Lectin (LTL) (Vector Laboratories, cat. no. B-1325) or biotinylated Dolichos biflorus Agglutinin (DBA) (Vector Laboratories, ca. no. B-1035) and Cellection Biotin Binder kit (ThermoFischer, cat. no. 11533D). Cells were immortalized using the large T antigen (Addgene plasmid no. 22298). Pkd1 was conditionally inactivated using cre recombinase (121112C-LTL cells; Excellgen, cat. no. EG-1001) or viral transduction (121112-C DBA and 94414-LTL/DBA cells) using LV-Cre (Addgene plasmid no. 12106). At the time of inactivation, a corresponding control was generated using viral transductionwith plasmid LV-Lac (Addgene plasmid no. 12108). Pkd1 inactivation was confirmed using genomic PCR and/or reverse-transcriptase PCR (TaqMan gene expression assay, Applied Biosystems, cat. no. 4351372, Mm00465436_g1). mCCDcl1 (mCCD) cells were a kind gift from the Rossier lab (Gaeggeler et al., 2005). mCCD Pkd1 knock-down cells were generated using viral transduction with the shRNA clone TRCN0000072085 and the corresponding pLKO.1 TRC21 control (Addgene plasmid 10,879). Cells were grown in DMEM/F12 media (Life cat. no. 21041–025) with 2% FBS (GEMINI Bio-Products cat. no. 100–106), 1X Insulin-Transferrin-Selenium (Thermo Fisher Scientific, cat. no. 41400–045), 5 uM dexamethasone (SIGMA, cat. no. D1756), 10 ng/ml EGF (SIGMA, cat. no. SRP3196), 1 nM 3,3′,5-Triiodo-L-thyronine (SIGMA, cat. no. T6397) and 10 mM HEPES (CORNING, cat. no. 25-060-CI). Mouse embryonic fibroblasts (MEFs) were obtained from E12.5 and E13.5 Pkd1 knockout (Pkd1tm1GGG (Bhunia et al., 2002)) and control mice. Briefly, whole embryos were minced, washed in PBS and cultured in six-well tissue culture plate in Dulbecco's minimal essential medium (DMEM) supplemented with 10% fetal bovine serum (FBS). A total of 6 MEF lines was used for this study: three from E13.5 mouse littermates (2 Pkd1ko/ko and 1 Pkd1wt/wt) immortalized using large T antigen (Addgene plasmid no. 22,298) and an additional set of three primary (passage 2, non-immortalized) E12.5 embryos (1 Pkd1ko/ko and 2 Pkd1wt/wt littermate controls). |
| Sample Type: | Tissue |
